Due to its turbulent demographic history, marked by extensive settlement and gene flow from diverse regions of Eurasia, Southeastern Europe (SEE) has consistently served as a genetic crossroads between East and West and a junction for the migrations that reshaped Europe’s population. SEE, including modern Croatian territory, was a crucial passage from the Near East and even more distant regions and human populations in this region, as almost any other European population represents a remarkable genetic mixture. Modern humans have continuously occupied this region since the Upper Paleolithic era, and different (pre)historical events have left a distinctive genetic signature on the historical narrative of this region. Our views of its history have been mostly renewed in the last few decades by extraordinary data obtained from Y-chromosome studies. In recent times, the international research community, bringing together geneticists and archaeologists, has steadily released a growing number of ancient genomes from this region, shedding more light on its complex past population dynamics and shaping the genetic pool in Croatia and this part of Europe.

Over the past 30 years, forensic experts from Croatia and Bosnia and Herzegovina have embraced advanced technologies and innovations to enable great efficacy and proficiency in the identification of war victims. The wartime events in the countries of former Yugoslavia greatly influenced the application of the selected DNA analyses as routine tools for the identification of skeletal remains, especially those from mass graves. Initially, the work was challenging because of the magnitude of the events, technical aspects, and political aspects. Collaboration with reputable foreign forensic experts helped tremendously in the efforts to start applying DNA analysis routinely and with increasing success. In this article, we reviewed the most significant achievements related to the application of DNA analysis in identifying skeletal remains in situations where standard identification methods were insufficient.

Aim To assess the association of single nucleotide polymorphisms (SNPs) in the ACE2 and TMPRSS2 genes with COVID-19 severity and key biomarkers. Methods The study involved 750 COVID-19 patients from Bosnia and Herzegovina, divided into three groups: mild, moderate, and severe cases. Genetic variations within the ACE2 (rs2285666) and TMPRSS2 (rs2070788) genes were examined with real-time polymerase chain reaction. Biochemical markers were determined with standard procedures. Results There was a significant difference in the rs2070788 genotype distribution between patients with mild and moderate symptoms, but not between other groups. For the rs2285666 polymorphism, no significant difference in genotype distribution was found. In patients with mild symptoms, carriers of the GG genotype of rs2070788 had significantly higher total bilirubin levels than carriers of the AA genotype. Similarly, carriers of the TT genotype of rs2285666 had significantly higher activated partial thromboplastin time and international normalized ratio, and lower lactate dehydrogenase levels compared with the CC genotype. Among patients with severe symptoms, carriers of the GG genotype showed significantly higher potassium levels than carriers of the AA genotype, while carriers of the TT genotype showed significantly higher erythrocyte count as well as hemoglobin and hematocrit levels compared with the CC genotype. Conclusion This study highlights the role of genetic factors, particularly SNPs in the ACE2 and TMPRSS2 genes, in determining COVID-19 severity, aiding patient risk assessment and prognosis.

Introduction: COVID-19 has been a major focus of scientific research since early 2020. Due to its societal, economic, and clinical impact worldwide, research efforts aimed, among other questions, to address the effect of host genetics in susceptibility and severity of COVID-19. Methods: We, therefore, performed next-generation sequencing of coding and regulatory regions of 16 human genes, involved in maintenance of the immune system or encoding receptors for viral entry into the host cells, in a subset of 60 COVID-19 patients from the General Hospital Tešanj, Bosnia and Herzegovina, classified into three groups of clinical conditions of different severity (“mild,” “moderate,” and “severe”). Results: We confirmed that the male sex and older age are risk factors for severe clinical picture and identified 13 variants on seven genes (CD55, IL1B, IL4, IRF7, DDX58, TMPRSS2, and ACE2) with potential functional significance, either as genetic markers of modulated susceptibility to SARS-CoV-2 infection or modifiers of the infection severity. Our results include variants reported for the first time as potentially associated with COVID-19, but further research and larger patient cohorts are required to confirm their effect. Discussion: Such studies, focused on candidate genes and/or variants, have a potential to answer the questions regarding the effect of human genetic makeup on the expected infection outcome. In addition, loci we identified here were previously reported to have clinical significance in other diseases and viral infections, thus confirming a general, broader significance of COVID-19-related research results following the end of the pandemic period.

Aim Examination of the effectiveness of STR loci in proving sibship of the Bosnian-Herzegovinian village of Orahovica and the formation of a "grey zone". Methods The probability of sibship was determined by calculating the likelihood ratio (LR) parameter for each of the 15 observed STR loci and for each of the pairs of relatives and non-relatives. Cumulative sibship index (CSI) was calculated for each of the pairs by multiplying the LR values of all 15 loci and obtained values are used as CSI limit for separating relatives from non-relatives. By creating a grey zone for local populations, an attempt was made to obtain a line of demarcation between siblings and non-siblings. Results An analysis of the origin of the respondents' relatives was performed, up to the level of sibship in the third generation. The results of the CSI for pairs of relatives from the village of Orahovica showed that the highest CSI value, and therefore the sibship probability was recorded among relatives from the village of Orahovica (CSI=534211727.203;SP=99.999999812%). On the contrary, incredibly low CSI value was recorded among non-relatives,ranging from CSI=0.0000001 to 0.5261434 (SP=0.000009999% to 34.475357951%). Conclusion For the threshold value CSI=1 and for CSI=3, this method determined sibship in 100% of pairs of relatives and the absence of biological sibship in 100% of pairs of non-relatives in the village of Orahovica. The STR system is proved to be a successful method in determining sibship or absence of sibship in small local populations.

Mitochondrial DNA (mtDNA) has been used for decades as a predominant tool in population genetics and as a valuable addition to forensic genetic research, owing to its unique maternal inheritance pattern that enables the tracing of individuals along the maternal lineage across numerous generations. The dynamic interplay between evolutionary forces, primarily genetic drift, bottlenecks, and the founder effect, can exert significant influence on genetic profiles. Consequently, the Adriatic islands have accumulated a subset of lineages that exhibits remarkable absence or rarity within other European populations. This distinctive genetic composition underscores the islands’ potential as a significant resource in phylogenetic research, with implications reaching beyond regional boundaries to contribute to a global understanding. In the initial attempt to expand the mitochondrial forensic database of the Croatian population with haplotypes from small isolated communities, we sequenced mitogenomes of rare haplogroups from different Croatian island and mainland populations using next-generation sequencing (NGS). In the next step and based on the obtained results, we refined the global phylogeny of haplogroup N1a, HV2, and X by analyzing rare haplotypes, which are absent from the current phylogenetic tree. The trees were based on 16 novel and 52 previously published samples, revealing completely novel branches in the X and HV2 haplogroups and a new European cluster in the ancestral N1a variant, previously believed to be an exclusively African–Asian haplogroup. The research emphasizes the importance of investigating geographically isolated populations and their unique characteristics within a global context.

This study investigates the genetic diversity and relatedness among a small local population in Bosnia and Herzegovina. In a genetic research study, a sample of 38 individuals was collected from the village of Vukotići, consisting of 21 male and 17 female subjects. The total genomic DNA was extracted using a modified Miller protocol. The QUANTIFILER DNA identification kit was used to quantify the total human DNA in the sample. The sibship relationship was assessed by computing the likelihood ratio for each of the 15 STR loci in both relatives and non-relatives. Results showed a higher homogeneity of the small local population compared to the mixed population within the larger population. Variability in peak height observed in the genetic analysis was attributed to differences in DNA concentration in the extracted samples. Probability of relatedness among participants in the Vukotići village was found to be low. Central tendency and variability measures revealed valuable insights into sample distribution and variation. The study concludes that CSI=1 and CSI=3 can be used as reliable tools to determine sibship in small local populations without a "gray zone".

Introduction: Cheiloscopy is defined as the study of the sulci labiorum, known as "lip prints". Aim of the study: The aim of this study was to determine the morphological characteristics of the lip grooves in the three dominant nationalities in the Republic of North Macedonia (Macedonians, Albanians, Roma) and to compare the obtained results with the morphological characteristics of the lip grooves in three other populations from different geographical regions. Material and methods: In this research, we included 150 examinees aged 25-50 years and divided them into three groups: Macedonians (50), Albanians (50) and Roma (50). The lip prints were taken using microscopic slides and detected using the dactyloscopic powder and brush method. We used the Suzuki and Tsuchihashi classification to typify the lip prints. Results: The most common type of lip grooves in the population of the Republic of North Macedonia was the type II grooves. There was no significant difference in the presence of different types of lip grooves in the four quadrants between males and females, nor between Macedonians, Albanians and Roma. The comparative analysis showed that populations from different geographical areas had different anthropological and morphological characteristics of the lip grooves. Conclusion: Type II lip grooves are the most common in the population of the Republic of North Macedonia and there is no statistically significant difference between the prevalence of different types of lip grooves in the three nationalities in this study. Considering the large number of factors that can affect the quality of the lip print

Aim To analyze an additional set of Y-chromosome genetic markers to acquire a more detailed insight into the diversity of the Croatian population. Methods A total of 518 Yfiler Plus profiles were genotyped. Allele frequencies, haplotype frequencies, and haplotype diversity were calculated by using the STRAF software v. 2.0.4. Genetic distances were quantified by Rst with AMOVA online tool from the YHRD. The evolutionary history was inferred with the neighbor-joining method of phylogenetic tree construction in the MEGAX software. Whit Athey's Haplogroup Predictor v. 5 was used for additional comparison with regional and other European populations. Results A total of 507 haplotypes were used for genetic STR analysis. An interpopulation study on 17 Y-STR markers showed the lowest genetic diversity between the Croatian and Bosnian-Herzegovinian populations and the highest between the Croatian and Irish populations. Additional interpopulation comparison with the original 27 Y-STR markers (for the population with available data) was also performed. A total of 518 haplotypes were used in the determination of haplogroup diversity. Haplogroup I with its sublineage I2a expressed the highest prevalence. The second most prevalent haplogroup was R, with its major sublineage R1a, except for the subpopulation of Hvar, where E1b1b was the second most prevalent haplogroup. Rare haplogroups also confirmed in this study were L, T, and Q. G1 was detected for the first time in the Croatian population. Conclusion We obtained a new insight into the differences between examined subpopulations of Croatia and their possible (dis)similarities with neighboring and distant populations.

Aim To use the method of meta-analysis to assess the influence of island population isolation on the sub-structuring of the Croatian population, as well as the influence of regional population groups on the sub-structuring of the Southeastern European population with regard to basic population genetic statistical parameters calculated by using STR locus analysis. Methods Bio-statistical analyses were performed for 2877 unrelated participants of both sexes from Southeastern Europe. Nine autosomal STR loci (D3S1358, vWA, FGA, TH01, TPOX, CSF1PO, D5S818, D13S317, and D7S82) were analyzed by using standard F-statistics and population structure analysis (Structure software). Results Genetic differentiation of Croatian subpopulations assessed with the FST method was higher at the level of the Croatian population (0.005) than at the level of Southeastern Europe (0.002). The island of Vis showed the most pronounced separation in the Croatian population, and Albanians from Kosovo in the population of Southeast Europe, followed by Croatia, Bosnia and Herzegovina, and Hungary. Conclusion The higher structure of Croatian subpopulations in relation to Southeastern Europe suggest a certain degree of genetic isolation, most likely due to the influence of endogamy within rural island populations.

Aim To investigate the influence of specific intrapopulation genetic structures on interpopulation relationships. Special focus was the influence of island population isolation on the substructuring of the Croatian population, and the influence of regional population groups on the substructuring of Southeast European populations. Methods Autosomal short tandem repeat (STR) loci were analyzed by using four forensic parameters: matching probability (PM), power of discrimination (PD), power of exclusion (PE), and polymorphic information content (PIC) on a sample of 2877 unrelated participants of both sexes. A sample set comprising 590 participants was analyzed for the first time, and 2287 participants were included from previous studies. The analysis was performed with PowerStats v. 1.2. Results The analysis of forensic parameters for all nine loci in the Croatian subpopulations showed the largest deviations in the populations of the islands of Korčula and Hvar. The smallest deviations were found in the mainland population. As for Southeast European populations, the largest deviations were found in the population of North Macedonia, followed by Romania, Albanians from Kosovo, and Montenegro, while the smallest deviations were found in the population of Hungary. Conclusion The comparison of forensic parameters between different subpopulations of Croatia and Southeast Europe indicates that the isolation of individual Croatian subpopulations and rare alleles in their gene pool affect the values of forensic parameters. Specific features of (sub)populations should be taken into account for appropriate sampling of the total population when creating a DNA database of STR markers.

Background Serostudies are important resources when following pandemics and predicting their further spread, as well as determining the length of protection against reinfection and vaccine development. The aim of this study was to update data on the prevalence of seropositive individuals in Canton Sarajevo, Bosnia and Herzegovina (B&H) from September 2020 to May 2021. Methods Anti-SARS-CoV-2 antibodies were quantified using an electrochemiluminescence immunoassay. Results Compared to the period April–July 2020, when anti-SARS-CoV-2 antibodies were detected in 3.77% of samples, one year later (May 2021) the estimated percentage within the same population of the urban Canton Sarajevo was 29.9% (5,406/18,066). Of all anti-SARS-CoV-2 Ig-positive individuals, 53.27% were men, and 69.00% were of 50 years of age or younger. Also, the current update found the individuals 50 years of age or younger to be more frequently anti-SARS-CoV-2 Ig positive compared to older individuals. On the other hand, higher median anti-SARS-CoV-2 Ig levels were found in individuals > 50 years old than in younger individuals, as well as in men compared to women. Seropositivity gradually increased from September 2020 to May 2021, with the lowest frequency of positive cases (3.5%) observed in September 2020, and the highest frequency (77.7%) in January 2021. Conclusion Our results provided important seroprevalence data that could help in planning restrictive local public health measures to protect the population of Sarajevo Canton, especially considering that at the time of the study the vaccines were virtually inaccessible to the general population not belonging to any of the high-priority groups for vaccination.